CA2744153C - Compositions et procedes pour l'assemblage de polynucleotides - Google Patents

Compositions et procedes pour l'assemblage de polynucleotides Download PDF

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Publication number
CA2744153C
CA2744153C CA2744153A CA2744153A CA2744153C CA 2744153 C CA2744153 C CA 2744153C CA 2744153 A CA2744153 A CA 2744153A CA 2744153 A CA2744153 A CA 2744153A CA 2744153 C CA2744153 C CA 2744153C
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nucleic acid
annealable linker
linker sequence
sequence
acid molecules
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CA2744153A1 (fr
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Zach Serber
Raymond Lowe
Jeffrey A. Ubersax
Sunil S. Chandran
Erik Jedediah Dean
Darren M. Platt
Kenneth Toshiki Takeoka
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Amyris Inc
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Amyris Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms

Abstract

La présente invention concerne des compositions et des procédés pour lassemblage rapide dun ou plusieurs polynucléotides assemblés parmi une pluralité de polynucléotides composants. Les procédés de linvention utilisent des vecteurs dacide nucléique circulaires qui comprennent un segment dADN D flanqué par une séquence de lieur hybridable, des paires de séquences de lieur hybridables LA et LB, ou des paires de séquence de lieur hybridable / segment de liaison damorce LA et PB ou PA et LB. La digestion par des endonucléases de restriction dune pluralité de vecteurs contenant les segments dADN à assembler génère une pluralité de fragments dADN comprenant les éléments PA-D-LB, LA-D-LB, et LA-D-PB ou D-LB, LA-D-LB, et LA-D. Les séquences de séquences de lieur hybridables LA et LB confèrent des terminaisons complémentaires aux fragments dADN, qui sont utilisées dans la recombinaison homologue véhiculée par une cellule hôte ou conjointement avec des segments de liaison damorce PA et PB dans une réaction dassemblage par cyclage de polymérase pour lassemblage ordonné des différents segments dADN en un ou plusieurs polynucléotides assemblés.
CA2744153A 2008-11-19 2009-11-19 Compositions et procedes pour l'assemblage de polynucleotides Active CA2744153C (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US11610908P 2008-11-19 2008-11-19
US61/116,109 2008-11-19
US16223009P 2009-03-20 2009-03-20
US61/162,230 2009-03-20
PCT/US2009/065048 WO2010059763A1 (fr) 2008-11-19 2009-11-19 Compositions et procédés pour l’assemblage de polynucléotides

Publications (2)

Publication Number Publication Date
CA2744153A1 CA2744153A1 (fr) 2010-05-27
CA2744153C true CA2744153C (fr) 2020-07-07

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Family Applications (1)

Application Number Title Priority Date Filing Date
CA2744153A Active CA2744153C (fr) 2008-11-19 2009-11-19 Compositions et procedes pour l'assemblage de polynucleotides

Country Status (12)

Country Link
US (3) US8221982B2 (fr)
EP (1) EP2358875B1 (fr)
JP (1) JP2012509084A (fr)
KR (1) KR101794298B1 (fr)
CN (1) CN102282255B (fr)
AU (1) AU2009316660B2 (fr)
BR (1) BRPI0922187B1 (fr)
CA (1) CA2744153C (fr)
MX (1) MX2011005195A (fr)
SG (1) SG171760A1 (fr)
WO (1) WO2010059763A1 (fr)
ZA (1) ZA201103637B (fr)

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Also Published As

Publication number Publication date
US20120245056A1 (en) 2012-09-27
CN102282255A (zh) 2011-12-14
KR20110102351A (ko) 2011-09-16
WO2010059763A1 (fr) 2010-05-27
BRPI0922187B1 (pt) 2021-05-04
US20100124768A1 (en) 2010-05-20
CA2744153A1 (fr) 2010-05-27
SG171760A1 (en) 2011-07-28
ZA201103637B (en) 2012-08-29
EP2358875B1 (fr) 2015-08-12
US8221982B2 (en) 2012-07-17
CN102282255B (zh) 2014-05-07
JP2012509084A (ja) 2012-04-19
US20100136633A1 (en) 2010-06-03
AU2009316660B2 (en) 2015-01-29
BRPI0922187A2 (pt) 2018-10-23
KR101794298B1 (ko) 2017-11-20
US8546136B2 (en) 2013-10-01
AU2009316660A1 (en) 2011-07-07
MX2011005195A (es) 2011-09-01
EP2358875A1 (fr) 2011-08-24
US8110360B2 (en) 2012-02-07

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